The liver is a key endoderm-derived multifunctional organ within the digestive system.Prospero homeobox 1(Prox1)is an essential transcription factor for liver development,but its specific function is not well understo...The liver is a key endoderm-derived multifunctional organ within the digestive system.Prospero homeobox 1(Prox1)is an essential transcription factor for liver development,but its specific function is not well understood.Here,we show that hepatic development,including the formation of intrahepatic biliary and vascular networks,is severely disrupted in prox1a mutant zebrafish.We find that Prox1a is essential for liver growth and proper differentiation but not required for early hepatic cell fate specification.Intriguingly,prox1a depletion leads to ectopic initiation of a Cdx1b-mediated intestinal program and the formation of intestinal lumen-like structures within the liver.Morpholino knockdown of cdx1b alleviates liver defects in the prox1a mutant zebrafish.Finally,chromatin immunoprecipitation analysis reveals that Prox1a binds directly to the promoter region of cdx1b,thereby repressing its expression.Overall,our findings indicate that Prox1a is required to promote and protect hepatic development by repression of Cdx1b-mediated intestinal cell fate in zebrafish.展开更多
目的:探讨尾侧同源盒转录因子1(CDX1)对结直肠癌细胞增殖凋亡及p38MAPK磷酸化水平的影响。方法:用荧光定量PCR和Western blot检测正常肠上皮细胞FHC和结直肠癌细胞SW480、HCT116、Caco2中CDX1 m RNA和蛋白表达水平。在SW480细胞中转染p ...目的:探讨尾侧同源盒转录因子1(CDX1)对结直肠癌细胞增殖凋亡及p38MAPK磷酸化水平的影响。方法:用荧光定量PCR和Western blot检测正常肠上皮细胞FHC和结直肠癌细胞SW480、HCT116、Caco2中CDX1 m RNA和蛋白表达水平。在SW480细胞中转染p IRES-CDX1和p IRES记为CDX1组和载体组,以不做转染的细胞为对照组,荧光定量PCR和Western blot测定CDX1 m RNA和蛋白表达水平,MTT检测细胞增殖,流式细胞术检测细胞凋亡,Western blot测定细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)、信号转导与转录因子3(STAT3)、磷酸化的STAT3(p-STAT3)、p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化的p38MAPK(p-p38MAPK)蛋白水平。结果:结直肠癌细胞SW480、HCT116、Caco2中CDX1 m RNA和蛋白表达水平均明显低于正常肠上皮细胞FHC(P<0.05),SW480细胞中CDX1 m RNA和蛋白表达水平明显低于HCT116、Caco2(P<0.05)。CDX1组细胞中CDX1 m RNA和蛋白水平明显高于对照组(P<0.05),载体组细胞中CDX1 m RNA和蛋白水平与对照组相比没有统计学差异(P>0.05)。CDX1组细胞存活率明显降低,细胞凋亡率明显升高,细胞中Cleaved Caspase-3、p-p38MAPK水平升高,细胞中Bcl-2、p-STAT3水平降低,与对照组相比差异具有统计学意义(P<0.05)。载体组细胞存活率、凋亡率及细胞中Cleaved Caspase-3、Bcl-2、STAT3、p-STAT3、p38MAPK、pp38MAPK蛋白水平与对照组相比没有明显变化(P>0.05)。结论:CDX1抑制结直肠癌细胞增殖,诱导Caspase-3介导的细胞凋亡,促进细胞中p38MAPK磷酸化,抑制细胞中STAT3磷酸化。展开更多
In mammals,the expression of the homeobox family member Cdx2/CDX2 is restricted within the intestine.Conditional ablation of the mouse Cdx2 in the endodermal cells causes a homeotic transformation of the intestine tow...In mammals,the expression of the homeobox family member Cdx2/CDX2 is restricted within the intestine.Conditional ablation of the mouse Cdx2 in the endodermal cells causes a homeotic transformation of the intestine towards the esophagus or gastric fate.In this report,we show that null mutants of zebrafish cdx1b,encoding the counterpart of mammalian CDX2,could survive more than 10 days post fertilization,a stage when the zebrafish digestive system has been well developed.Through RNA sequencing(RNA-seq)and single-cell sequencing(sc RNA-seq)of the dissected intestine from the mutant embryos,we demonstrate that the loss-of-function of the zebrafish cdx1b yields hepatocyte-like intestinal cells,a phenotype never observed in the mouse model.Further RNA-seq data analysis,and genetic double mutants and signaling inhibitor studies reveal that Cdx1b functions to guard the intestinal fate by repressing,directly or indirectly,a range of transcriptional factors and signaling pathways for liver specification.Finally,we demonstrate that heat shock-induced overexpression of cdx1b in a transgenic fish abolishes the liver formation.Therefore,we demonstrate that Cdx1b is a key repressor of hepatic fate during the intestine specification in zebrafish.展开更多
基金partially supported by grants from the National Key Research and Development Program of China and the National Natural Science Foundation of China(2018YFA0801000,32270889,2019YFA0802800,32070824,2015CB942800,2016YFA0100500,31871458,31671500,and 81371264)supported by Beijing Natural Science Foundation(5242009)a grant from the Fisheries Innovation Team of Beijing Agriculture Innovation Consortium(BAIC07-2023-02).
文摘The liver is a key endoderm-derived multifunctional organ within the digestive system.Prospero homeobox 1(Prox1)is an essential transcription factor for liver development,but its specific function is not well understood.Here,we show that hepatic development,including the formation of intrahepatic biliary and vascular networks,is severely disrupted in prox1a mutant zebrafish.We find that Prox1a is essential for liver growth and proper differentiation but not required for early hepatic cell fate specification.Intriguingly,prox1a depletion leads to ectopic initiation of a Cdx1b-mediated intestinal program and the formation of intestinal lumen-like structures within the liver.Morpholino knockdown of cdx1b alleviates liver defects in the prox1a mutant zebrafish.Finally,chromatin immunoprecipitation analysis reveals that Prox1a binds directly to the promoter region of cdx1b,thereby repressing its expression.Overall,our findings indicate that Prox1a is required to promote and protect hepatic development by repression of Cdx1b-mediated intestinal cell fate in zebrafish.
文摘目的:探讨尾侧同源盒转录因子1(CDX1)对结直肠癌细胞增殖凋亡及p38MAPK磷酸化水平的影响。方法:用荧光定量PCR和Western blot检测正常肠上皮细胞FHC和结直肠癌细胞SW480、HCT116、Caco2中CDX1 m RNA和蛋白表达水平。在SW480细胞中转染p IRES-CDX1和p IRES记为CDX1组和载体组,以不做转染的细胞为对照组,荧光定量PCR和Western blot测定CDX1 m RNA和蛋白表达水平,MTT检测细胞增殖,流式细胞术检测细胞凋亡,Western blot测定细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)、信号转导与转录因子3(STAT3)、磷酸化的STAT3(p-STAT3)、p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化的p38MAPK(p-p38MAPK)蛋白水平。结果:结直肠癌细胞SW480、HCT116、Caco2中CDX1 m RNA和蛋白表达水平均明显低于正常肠上皮细胞FHC(P<0.05),SW480细胞中CDX1 m RNA和蛋白表达水平明显低于HCT116、Caco2(P<0.05)。CDX1组细胞中CDX1 m RNA和蛋白水平明显高于对照组(P<0.05),载体组细胞中CDX1 m RNA和蛋白水平与对照组相比没有统计学差异(P>0.05)。CDX1组细胞存活率明显降低,细胞凋亡率明显升高,细胞中Cleaved Caspase-3、p-p38MAPK水平升高,细胞中Bcl-2、p-STAT3水平降低,与对照组相比差异具有统计学意义(P<0.05)。载体组细胞存活率、凋亡率及细胞中Cleaved Caspase-3、Bcl-2、STAT3、p-STAT3、p38MAPK、pp38MAPK蛋白水平与对照组相比没有明显变化(P>0.05)。结论:CDX1抑制结直肠癌细胞增殖,诱导Caspase-3介导的细胞凋亡,促进细胞中p38MAPK磷酸化,抑制细胞中STAT3磷酸化。
基金supported by Centre for Computational Science and Engineering(CCSE)at Southern University of Science and Technologysupported by the National Key R&D Program of China(2018YFA0800502)the National Natural Science Foundation of China(31900579,31830113)。
文摘In mammals,the expression of the homeobox family member Cdx2/CDX2 is restricted within the intestine.Conditional ablation of the mouse Cdx2 in the endodermal cells causes a homeotic transformation of the intestine towards the esophagus or gastric fate.In this report,we show that null mutants of zebrafish cdx1b,encoding the counterpart of mammalian CDX2,could survive more than 10 days post fertilization,a stage when the zebrafish digestive system has been well developed.Through RNA sequencing(RNA-seq)and single-cell sequencing(sc RNA-seq)of the dissected intestine from the mutant embryos,we demonstrate that the loss-of-function of the zebrafish cdx1b yields hepatocyte-like intestinal cells,a phenotype never observed in the mouse model.Further RNA-seq data analysis,and genetic double mutants and signaling inhibitor studies reveal that Cdx1b functions to guard the intestinal fate by repressing,directly or indirectly,a range of transcriptional factors and signaling pathways for liver specification.Finally,we demonstrate that heat shock-induced overexpression of cdx1b in a transgenic fish abolishes the liver formation.Therefore,we demonstrate that Cdx1b is a key repressor of hepatic fate during the intestine specification in zebrafish.
